S-Space College of Medicine/School of Medicine (의과대학/대학원) Dept. of Medicine (의학과) Theses (Ph.D. / Sc.D._의학과)
Functional study of recombinant Clonorchis sinensis thioredoxin peroxidase on inhibitory effect of macrophage activation
재조합한 간흡충 thioredoxin peroxidase의 대식세포 기능 억제 효능 연구
- 의과대학 의학과
- Issue Date
- 서울대학교 대학원
- Clonorchis sinensis; thioredoxin peroxidase; deletional mutant; site-directed mutagenesis; immune evasion; LPS; IL-10; TGF-β; nitric oxide; IL-6; macrophages
- 학위논문 (박사)-- 서울대학교 대학원 : 의학과, 2014. 8. 배영미.
- Clonorchis sinensis (C. sinensis) estabilishes long term parasitism within bile ducts of humans without notable symptoms. General down-modulated host immune responses in helminth infestation are maintained by excretory-secretory products (ESP), produced by the live parasites and considered as an important immunomodulator in this context. Co-culture experiments with RAW264.7 cells and C. sinensis adults worms through permeable transwell demonstrated that ESP from living C. sinensis functioned as a ligand for TLR4, but effectively attenuated LPS-induced IL-6 and NO production. On the other hands, upsurged secretion of antiinflammatory cytokines IL-10 and TGF-β were observed by C. sinensis ESP treatment. Reference base study found out that thioredoxin peroxidase (TPX) of helminth parasites had an immune-modulating property toward Th2 type through induction of regulatory phenotype of macrophages. Hence, recombinant TPX of C. sinensis was produced and its gene was registered at Genbank (AND65138.1). The CS-TPX reduced LPS-induced expression of TLR4 on cell surface of RAW264.7 cells and LPS-induced secretion of pro-inflammatory mediators NO, TNF-α and IL-6. Moreover, CS-TPX induces IL-10, but not TGF-β. Pull-down assay result revealed the direct interaction between CS-TPX and TLR4 protein. Furthermore, the smallest recombinant fragment of CS-TPX, which is T44, has comparable anti-inflammatory. In conclusion, this study described the macrophage regulatory function of C. sinensis ESP and the smallest fragment of CS-TPX, T44, could function of the negative regulator in LPS treated macrophage cell line RAW264.7 cells. Defined CS-TPX T44 will be one of the best candidates for therapeutic immunosuppressant in allergies and autoimmune diseases.